实时荧光定量PCR检测高温即时浸酸对蚕卵中家蚕微孢子虫的灭活效果
|
摘要
家蚕微孢子虫(Nosema bombycis,Nb)引起的家蚕微粒子病是对蚕种生产危害最为严重的家蚕病害,蚕种浸酸处理是蚕业生产中常用的一种防病处理手段。为探明浸酸处理对家蚕微粒子病的防治效果,将家蚕二化性四元杂交组合两广二号的带毒蚕卵分别进行常规即时浸酸处理和高温即时浸酸处理,采用实时荧光定量PCR对卵龄24～240 h蚕卵中的家蚕微孢子虫进行定量检测。结果发现,常规和高温浸酸处理后每日取样蚕卵中家蚕微孢子虫的检测值较阳性对照组均显著降低,高温浸酸组又明显低于常规浸酸组,表明2种浸酸处理对蚕卵中的家蚕微孢子虫均有灭活作用,且高温处理的灭活效果更为显著。对蚕卵孵化后饲养至3龄眠和5龄起的幼虫逐条进行显微镜镜检,结果显示高温即时浸酸处理对家蚕微粒子病的相对防治效果分别为92.30%±2.95%和81.80%±0.06%。综上表明,高温即时浸酸处理带毒蚕卵对家蚕微粒子病有显著的防治作用。
Pebrine disease caused by Nosema bombycis(Nb) is the most serious silkworm disease in silkworm egg production, and acid treatment is a kind of commonly used treatment to control silkworm disease. To improve the control effect of acid treatment on pebrine disease, conventional and high temperature instant acid treatment were carried out on silkworm eggs of the bivoltine quaternary hybrid combination Liangguang 2. Then real-time fluorescence quantitative PCR was used to detect Nb in eggs 24 to 240 h after laying. Results showed that the detected value of Nb in both conventional and high temperature instant acid treated silkworm eggs was significantly lower than that in positive control group, and the inactivation effect of high temperature instant acid treatment was better than that of the conventional acid treatment, which indicated that acid treatments can inactivate the pathogens in silkworm eggs and high temperature can enhance the effect. Moreover, when examining silkworm larvae hatched from treated eggs in the 3 rd and 5 th instar under microscopy, we found that the relative control effect of high temperature instant acid treatment on pebrine disease reached 92.30%±2.95% and 81.80%±0.06% respectively. In summary, high temperature instant acid treatment to infected silkworm eggs is an effective way to prevent pebrine disease in silkworm.
Pebrine disease caused by Nosema bombycis(Nb) is the most serious silkworm disease in silkworm egg production, and acid treatment is a kind of commonly used treatment to control silkworm disease. To improve the control effect of acid treatment on pebrine disease, conventional and high temperature instant acid treatment were carried out on silkworm eggs of the bivoltine quaternary hybrid combination Liangguang 2. Then real-time fluorescence quantitative PCR was used to detect Nb in eggs 24 to 240 h after laying. Results showed that the detected value of Nb in both conventional and high temperature instant acid treated silkworm eggs was significantly lower than that in positive control group, and the inactivation effect of high temperature instant acid treatment was better than that of the conventional acid treatment, which indicated that acid treatments can inactivate the pathogens in silkworm eggs and high temperature can enhance the effect. Moreover, when examining silkworm larvae hatched from treated eggs in the 3 rd and 5 th instar under microscopy, we found that the relative control effect of high temperature instant acid treatment on pebrine disease reached 92.30%±2.95% and 81.80%±0.06% respectively. In summary, high temperature instant acid treatment to infected silkworm eggs is an effective way to prevent pebrine disease in silkworm.
引文
[1]周泽扬,潘国庆,向仲怀.家蚕微孢子虫研究10年回眸[J].蚕业科学,2014,40(6):949-956
[2]鲁兴萌,邵勇奇.家蚕微粒子病防控技术研究的发展现状与趋势[J].蚕业科学,2016,42(6):945-952
[3]廖森泰,杨琼,邢东旭,等.家蚕微粒子病全程防控技术体系简述[J].蚕业科学,2016,42(1):148-151
[4]陈启沅.蚕桑谱[M].桂林:广西师范大学出版社,2015:5-40
[5]阿华尼谢T T,拉勃柴尼特齐ВH,钱品娟.对微粒子病蚕卵采用短时加温浸汤消毒试验的初步结果[J].广东蚕丝通讯,1959,1(6):27-28
[6]黄旭华,朱方容,潘志新,等.温度对家蚕微孢子虫(Nosema bombycis)孢子体外发芽的影响[J].南方农业学报,2013,44(6):1026-1029
[7]邢东旭,肖阳,廖森泰,等.蚕卵高温即时浸酸阻断家蚕微粒子病胚种垂直传播的效果[J].蚕业科学,2018,44(1):100-104
[8]YANG Q,XING D X,LI R,et al.Standard method for detecting Bombyx mori nucleopolyhedrovirus disease-resistant silkworm varieties[J].Rev Bras Entomol,2018,62(1):19-22
[9]段廷云,陈红英,崔保安,等.实时荧光定量PCR检测H1N1亚型猪流感病毒[J].畜牧兽医学报,2008,39(6):752-756
[10]BUSTIN S A,BENES V,GARSON J A,et al.The MIQEguidelines:minimum information for publication of quantitative real-time PCR experiments[J].Clin Chem,2009,55(4):611-622
[11]邢东旭,肖阳,廖森泰,等.蚕卵高温即时浸酸阻断家蚕微粒子病胚种垂直传播的效果[J].蚕业科学,2018,44(1):100-104
[12]ISHIHARA R.The life cycle of Nosema bombycis as revealed in tissue culture cells of Bombyx mori[J].J Invertebr Pathol,1969,14(3):316-320
[13]邹宇晓.干热空气处理蚕卵防治微粒子病经卵传染的研究[D].广州:华南农业大学,1998
[14]王璐,杨琼,邢东旭,等.热处理法防治家蚕微粒子病的研究进展[J].广东蚕业,2018,52(2):1-3
[2]鲁兴萌,邵勇奇.家蚕微粒子病防控技术研究的发展现状与趋势[J].蚕业科学,2016,42(6):945-952
[3]廖森泰,杨琼,邢东旭,等.家蚕微粒子病全程防控技术体系简述[J].蚕业科学,2016,42(1):148-151
[4]陈启沅.蚕桑谱[M].桂林:广西师范大学出版社,2015:5-40
[5]阿华尼谢T T,拉勃柴尼特齐ВH,钱品娟.对微粒子病蚕卵采用短时加温浸汤消毒试验的初步结果[J].广东蚕丝通讯,1959,1(6):27-28
[6]黄旭华,朱方容,潘志新,等.温度对家蚕微孢子虫(Nosema bombycis)孢子体外发芽的影响[J].南方农业学报,2013,44(6):1026-1029
[7]邢东旭,肖阳,廖森泰,等.蚕卵高温即时浸酸阻断家蚕微粒子病胚种垂直传播的效果[J].蚕业科学,2018,44(1):100-104
[8]YANG Q,XING D X,LI R,et al.Standard method for detecting Bombyx mori nucleopolyhedrovirus disease-resistant silkworm varieties[J].Rev Bras Entomol,2018,62(1):19-22
[9]段廷云,陈红英,崔保安,等.实时荧光定量PCR检测H1N1亚型猪流感病毒[J].畜牧兽医学报,2008,39(6):752-756
[10]BUSTIN S A,BENES V,GARSON J A,et al.The MIQEguidelines:minimum information for publication of quantitative real-time PCR experiments[J].Clin Chem,2009,55(4):611-622
[11]邢东旭,肖阳,廖森泰,等.蚕卵高温即时浸酸阻断家蚕微粒子病胚种垂直传播的效果[J].蚕业科学,2018,44(1):100-104
[12]ISHIHARA R.The life cycle of Nosema bombycis as revealed in tissue culture cells of Bombyx mori[J].J Invertebr Pathol,1969,14(3):316-320
[13]邹宇晓.干热空气处理蚕卵防治微粒子病经卵传染的研究[D].广州:华南农业大学,1998
[14]王璐,杨琼,邢东旭,等.热处理法防治家蚕微粒子病的研究进展[J].广东蚕业,2018,52(2):1-3